NF-kappaB p65 (Phospho-S536) Rabbit Polyclonal Antibody

Datasheet

MSDS

NF-kappaB p65 (Phospho-S536) Rabbit Polyclonal Antibody

Key features and details

Rabbit polyclonal antibody to NF-kappaB p65 (Phospho-S536)

Target: NF-kappaB p65 (Phospho-S536)
Source/Host: Rabbit
Reactivity: Human,Mouse,Rat,Monkey,Pig
Clonality: Polyclonal
Applications: WB, IHC, IF/ICC, IP
Conjugation: Unconjugated
Storage: at-20°C

Brand:

CAT.NO. : APA07495

US$ Please choose

Size:

50μL 100μL

Trail, Bulk size or Custom requests Please contact us

Product Details

Background

NF-kappa-B is a pleiotropic transcription factor present in almost all cell types and is the endpoint of a series of signal transduction events that are initiated by a vast array of stimuli related to many biological processes such as inflammation, immunity, differentiation, cell growth, tumorigenesis and apoptosis. NF-kappa-B is a homo- or heterodimeric complex formed by the Rel-like domain-containing proteins RELA/p65, RELB, NFKB1/p105, NFKB1/p50, REL and NFKB2/p52. The heterodimeric RELA-NFKB1 complex appears to be most abundant one. The dimers bind at kappa-B sites in the DNA of their target genes and the individual dimers have distinct preferences for different kappa-B sites that they can bind with distinguishable affinity and specificity.

Application

To ensure optimal assay performance, AREX recommends conducting reagent titration tailored to each testing system for optimal detection results.

WB	1:500 - 1:1000
IHC	1:50 - 1:200
IF/ICC	1:50 - 1:200
IP	1:10 - 1:100

*Results are sample-specific. Please refer to your local assay conditions and test parameters for reference.

Overview

Description	Rabbit polyclonal antibody to NF-kappaB p65 (Phospho-S536)
Specificity	Recognizes endogenous levels of NF-kappaB p65 protein only when phosphorylated at S536.
Antibody Type	Primary antibody
Imnunogen	KLH-conjugated synthetic phosphopeptide corresponding to residues surrounding S536 of human NF-kappaB p65 protein. The exact sequence is proprietary.
Purification	The antibody was purified by immunogen affinity chromatography.
Molecular Weight	Predicted: 60 kD; Observed: 75 kD
Form/Buffer	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.01% sodium azide.
Alternative Names	NFKB3; Transcription factor p65; Nuclear factor NF-kappa-B p65 subunit; Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3
Gene Symbol	RELA
Entrez Gene	5970(Human); 19697(Mouse)
SwissProt	Q04206(Human); Q04207(Mouse)

*AREX continuously optimizes our products. Webpage content may not reflect the latest updates. For inquiries, please contact info@arexbio.com or your local distributor.
*Clone Number, Reactivity, Source/Host and Clonality can be found in the product name and Key Features section above.

Data

Western blot analysis of NF-kappaB p65 (Phospho-S536) expression in Hela (A), HEK293T (B), NIH3T3 (C), mouse kidney (D), H9C2 (E) whole cell lysates. (Predicted band size: 60 kD; Observed band size: 75 kD)

Immunohistochemical analysis of NF-kappaB p65 (Phospho-S536) staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Immunofluorescent analysis of NF-kappaB p65 (Phospho-S536) staining in rat lung. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using a AREX® Fluor 594-conjugated secondary antibody (red) in PBS at room temperature in the dark. DAPI was used to stain the cell nuclei (blue).

Direct ELISA antibody dose-response curve using Anti-NF-kappaB p65 (Phospho-S536) Antibody. Antigen (Phosphopeptide and non-phosphopeptide) concentration is 5 ug/ml. Goat Anti-Rabbit IgG (H&L) - HRP was used as the secondary antibody, and signal was developed by TMB substrate.

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

Note

For Research Use Only. Not for diagnostic, therapeutics, prophylactic or in vivo use.

FAQs

What are the main types of research antibodies and how do they differ?

Research antibodies are mainly divided into monoclonal antibodies and polyclonal antibodies. Monoclonal antibodies typically offer higher specificity and better batch-to-batch consistency, while polyclonal antibodies often provide stronger affinity but may show more variation between batches. The choice depends on your specific experimental needs.

How can I tell if a research antibody is suitable for my experiment?

It is recommended to carefully review the product datasheet for validated applications, species reactivity, recommended dilutions, and published references. For new antibodies, performing a small-scale validation with positive control samples is usually helpful.

Can improper storage of research antibodies affect experimental results?

Yes. Antibodies are sensitive to temperature, repeated freeze-thaw cycles, and contamination. Improper storage may lead to reduced activity, increased background, or weaker signals. It is best to follow the storage instructions provided in the product datasheet.

Why doesn’t the recommended dilution in the datasheet work well in my experiment?

The recommended dilution is based on the supplier’s test conditions. Factors such as sample type, fixation method, and detection system in your lab can influence the optimal working concentration. Performing a dilution series optimization in your own system is often necessary.

What precautions should I take when using a newly purchased research antibody for the first time?

It is advisable to briefly centrifuge the antibody (especially concentrated or lyophilized ones), then perform a small-scale pilot experiment using the recommended conditions. Recording the batch number and usage date is also helpful for future tracking.

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